Bioengineering

Abstract. This paper proposes an ordering of ideas to create politics that distinguishes itself from conventional politics. It is called extrapolitics. To define extrapolitics, the concept of politics, transhumanism and dialectic - dialogism have been briefly included. The  Politics Path has been schematized as a process of evolution that politics experiments and goes hand in hand with human evolution. A post-politicist proposal was suggested for the proper proliferation of transhumanist objectives, as well as an extension of the conventional political spectrum, one that includes the extra-political spectrum and arguments against human enhancement. It finalizes with the inclusion of two potential phenomena, one harmful and the other one ideal, that define the processes of application of extrapolitical policies, the bio-socio-political singularity and the bio-socio-political inflection, respectively.

Key words: transhumanism, politics, philosophy, extrapolitics, dialogism, ideologies, singularity, postmodernism, complexity.

The liver is a key organ that can communicate with many other districts of the human body. In the last few decades, much interest has focused on the interaction between the liver and the gut microbiota, with their reciprocal influence on biosynthesis pathways and the integrity the intestinal epithelial barrier. Dysbiosis or liver disorders lead to0 epithelial barrier dysfunction, altering membrane permeability to toxins. Clinical and experimental evidence shows that the permeability hence the delivery of neurotoxins such as LPS, ammonia and salsolinol contribute to neurological disorders. These findings suggested multi-organ communication between the gut microbiota, the liver and the brain. With a view to in vitro modeling this liver-based multi-organ communication, we describe the latest advanced liver-on-a-chip devices and discuss the need for new organ-on-a-chip platforms for in vitro modeling the in vivo multi-organ connection pathways in physiological and pathological situations.

We are currently witnessing a major epidemic caused by the 2019 novel coronavirus (2019-nCoV). The evolution of 2019-nCoV remains elusive. We found 4 insertions in the spike glycoprotein (S) which are unique to the 2019-nCoV and are not present in other coronaviruses. Importantly, amino acid residues in all the 4 inserts have identity or similarity to those in the HIV-1 gp120 or HIV-1 Gag. Interestingly, despite the inserts being discontinuous on the primary amino acid sequence, 3D-modelling of the 2019-nCoV suggests that they converge to constitute the receptor binding site. The finding of 4 unique inserts in the 2019-nCoV, all of which have identity /similarity to amino acid residues in key structural proteins of HIV-1 is unlikely to be fortuitous in nature. This work provides yet unknown insights on 2019-nCoV and sheds light on the evolution and pathogenicity of this virus with important implications for diagnosis of this virus.

Solid-state fermentation (SSF) has built up credibility in recent years in biotech industries due to its potential applications in the production of biologically active secondary metabolites, apart from feed, fuel, food, industrial chemicals and pharmaceutical products and has emerged as an attractive alternative to submerged fermentation. Bioremediation, bioleaching, biopulping, biobeneficiation, etc. are the major applications of SSF in bioprocesses which have set another milestone. Utilization of agro-industrial residues as substrates in SSF processes provides an alternative avenue and value-addition to these otherwise under-or non-utilized residues. Innovation is the key to success and it is imperative to be up-to-date with the changing demands of the industries and meet their needs for better product and services. Better understanding of biochemical engineering aspects, particularly on mathematical modeling and design of bioreactors (fermenters) has made it possible to scale-up SSF processes and some designs have been developed for commercialization, making the technology economically feasible. In future, SSF technology would be well developed at par with SmF if rationalization and standardization continues in current trend. This review describes the state-of-art scenario in totality on SSF although the focus is on the most recent developments of last 5 years or so on SSF processes and products developments.

The antimicrobial effects of sliver ions have been known for long but their activity against microorganisms has not yet been conclusively studied. In the present study, biological synthesis of silver nanoparticles using Klebsiella pneumonia has been investigated. The bacterial strain was grown the appropriate production media. The cell free filtrate obtained on filtration was bought in contact with different concentrations of AgNO 3. . The change in the pH with respect to the different concentrations was studied .The nanoparticles were further characterized by SEM studies. The antimicrobial activity of silver nanoparticles was investigated against E. coli, Bacillus and ESBL .Also the antimicrobial activity of encapsulated beads of silver nanoparticles and 2mM silver nanoparticles was compared. The anti-microbial fabrics activity was studied by coating the silver nanoparticles on gauge cotton fabric and then compared with standard disc Oxacillin. UV-VIS studies showed an increased in absorbance at 363 nm. The antimicrobial tests against E.coli showed increased inhibition at 1mM while for Bacillus, inhibition was more prominent at 2mM concentration and for ESBL-Pseudomonas inhibitory concentration was 2mM .The zone of inhibition for the encapsulated beads was less than that of the 2mM silver nanoparticles.

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The design of the supply chain, and in particular of the distribution phase, for fresh-food products, such as fresh and fresh-cut produce, fruit or meat, cannot be achieved without considering the perishable nature and the variability of the products entering the chain.

Background: The use of embryos in embryonic stem cell research (ESCR) has elicited ethical controversies as it entails the destruction of 5-day old human embryos to harvest stem cells. Objective: To explore the ethical positions of Islam, Buddhism, Hinduism, and Catholicism concerning the use of (1) left-over embryos from in vitro fertilization (IVF) also known as ‘surplus’ embryos and (2) ‘research embryos’ which are created by scientists to conduct research using embryonic stem cells. Methods: The opinions of religious leaders of Buddhist, Hindu, and Catholic faiths in Malaysia pertaining to ESCR were examined via in-depth, semi-structured interviews while Islamic responses are collected from local writings related to the derivation of fatwa on this issue. Participants’ responses on the ethics of human stem cell research are presented as a reflection of various scriptural texts of these four religions. These are presented and supported with the help of international bioethics lite...

Hip arthroplasty has proven to be a reliable and suitable surgical procedure to return patients back to function. Patient selection, surgical procedures and of the condition and the use of an appropriate implant plays a tremendous role for the success of the surgery, without the need of a revision in the near time. [1] The design and the final inspection of these implants have a very important place in the whole procedure of their manufacturing, and of course in their tendency to failure. [2] For this reason strict international standards were instituted to protect and make reliable the design and the machining of them. In both these procedures the dimensional and geometrical accuracy are very important and the limits of the acceptance of any inaccuracy are very narrow. In this paper it will be presented the whole procedure of producing such parts, using specific CAD/ CAE / CAM software for that, and the inspection of the implants, femoral heads which were made, with the use of AFM for its surface roughness accuracy.

Biosynthetic approach using an endophytic fungus is a novel way towards the development of safe, economically viable and green method for the synthesis of silver nanoparticles and thus synthesized silver nanoparticles can be used in antibacterial formulations. We report the extracellular synthesis of silver nanoparticles using an fungus F.solani sp, and their antibacterial activity against human pathogenic bacteria viz. Staphylococcus aureus, Bacillus subtilis ,Escheriachia.coli and Salmonella typhi. Detection of synthesized silver nanoparticles was carried out using UV-Visible spectrophotometer analysis, which showed a peak ranging between 420-450 nm indicating the formation of nanoparticles.

Laccase gene IV from Trametes versicolor was inserted by REMI in Aspergillus niger C28B25. One selected transformant (C28eco3) was subjected to a second round of transformation and a new transformant was isolated (C28eco3-13). Laccase production was compared for these strains in submerged fermentation (SmF) and solid-state fermentation (SSF) using glucose or maltodextrin at different concentrations (10, 50 and 100 g/L). Enzymes yields were higher with glucose as a carbon source. However, repression of laccase was evident for SmF after increasing glucose from 10 to 50 g/L. In contrast, in SSF glucose at 50 g/L allowed the production of the highest laccase activity for strain C28eco3 (362 U/L) and C28eco3-13 (592 U/L). Advantages of SSF as an expression and production system over SmF are also discussed.

Culture-independent studies rely on the quantity and quality of the extracted environmental metagenomic DNA (mDNA). To fully access the plant tissue microbiome, the extracted plant mDNA should allow optimal PCR applications and the genetic content must be representative of the total microbial diversity. In this study, we evaluated the endophytic bacterial diversity retrieved using different mDNA extraction procedures. Metagenomic DNA from sorghum (Sorghum bicolor L. Moench) stem and root tissues were extracted using two classical DNA extraction protocols (CTAB-and SDS-based) and five commercial kits. The mDNA yields and quality as well as the reproducibility were compared. 16S rRNA gene terminal restriction fragment length polymorphism (t-RFLP) was used to assess the impact on endophytic bacterial community structures observed. Generally, the classical protocols obtained high mDNA yields from sorghum tissues; however, they were less reproducible than the commercial kits. Commercial kits retrieved higher quality mDNA, but with lower endophytic bacterial diversities compared to classical protocols. The SDS-based protocol enabled access to the highest sorghum endophytic diversities. Therefore, "SDS-extracted" sorghum root and stem microbiome diversities were analysed via 454 pyrosequencing, and this revealed that the two tissues harbour significantly different endophytic communities. Nevertheless, both communities are dominated by agriculturally important genera such as Microbacterium, Agrobacterium, Sphingobacterium, Herbaspirillum, Erwinia, Pseudomonas and Stenotrophomonas; which have previously been shown to play a role in plant growth promotion. This study shows that DNA extraction protocols introduce biases in culture-independent studies of environmental microbial communities by influencing the mDNA quality, which impacts the microbial diversity analyses and evaluation. Using the broad-spectrum SDSbased DNA extraction protocol allows the recovery of the most diverse endophytic communities associated with sorghum tissues and, as such, establishes a reliable basis for future study of endophytic communities.

A high throughput screening (HTS) strategy was investigated for screening high-yielding cephalosporin C (CPC) using mutated Cephalosporium acremonium. The strategy was based on microtiter plates (MTPs) to culture the microorganisms and measure its products. The feasibility of MTPs was to replace the cultivation in shake flasks, and CPC production will be determined using turbidimetric assay by ELISA reader to replace HPLC assay. The HTS systems were verified by screening high-producing CPC mutants. By using this HTS strategy, two mutants, i.e. 5-A1 and 8-A2 were selected from the secondary mutagenesis. The volumetric productivity of Cephalosporin C from Cephalosporium acremonium was 7914.43 U/ml; 8639.24 U/ml in shake flasks. CPC production was improved about 246% and 269% compared with that of original strain.

Precise control over scaffold material, porosity, and internal pore architecture is essential for tissue engineering. By coupling solid free form (SFF) manufacturing with conventional sponge scaffold fabrication procedures, we have developed methods for casting scaffolds that contain designed and controlled locally porous and globally porous internal architectures. These methods are compatible with numerous bioresorbable and non-resorbable polymers, ceramics, and biologic materials. Phase separation, emulsion-solvent diffusion, and porogen leaching were used to create poly(l)lactide (PLA) scaffolds containing both computationally designed global pores (500, 600, or 800 mm wide channels) and solvent fashioned local pores (50-100 mm wide voids or 5-10 mm length plates). Globally porous PLA and polyglycolide/PLA discrete composites were made using melt processing. Biphasic scaffolds with mechanically interdigitated PLA and sintered hydroxyapatite regions were fabricated with 500 and 600 mm wide global pores. PLA scaffolds with complex internal architectures that mimicked human trabecular bone were produced. Our indirect fabrication using casting in SFF molds provided enhanced control over scaffold shape, material, porosity and pore architecture, including size, geometry, orientation, branching, and interconnectivity. These scaffolds that contain concurrent local and global pores, discrete material regions, and biomimetic internal architectures may prove valuable for multi-tissue and structural tissue interface engineering. r

Microspheres are characteristically free flowing powders having particle size ranging from 1-1000 μm consisting of proteins or synthetic polymers. The range of Techniques for the preparation of microspheres offers a Variety of opportunities to control aspects of drug administration and enhance the therapeutic efficacy of a given drug. There are various approaches in delivering a therapeutic substance to the target site in a sustained controlled release fashion. It is the reliable means to deliver the drug to the target site with specificity, if modified, and to maintain the desired concentration at the site of interest. Moreover the microspheres are of micron size so they can easily fit into various capillary beds which are also having micron size. Microspheres received much attention not only for prolonged release, but also for targeting of anticancer drugs. There are various departments of medicine like cancer, pulmonary, cardiology, radiology, gynecology, and oncology etc., numerous drugs are used and they are delivered by various types of drug delivery system. Among them microspheric drug delivery system has gained enormous attention. The purpose of the review is to compile various types of microspheres, different methods to preparation, its applications and also various parameters to evaluate their efficiency.

Omega-hydroxyfatty acids are excellent monomers for synthesizing a unique family of polyethylene-like biobased plastics. However, ω-hydroxyfatty acids are difficult and expensive to prepare by traditional organic synthesis, precluding their use in commodity materials. Here we report the engineering of a strain of the diploid yeast Candida tropicalis to produce commercially viable yields of ω-hydroxyfatty acids. To develop the strain we identified and eliminated 16 genes encoding 6 cytochrome P450s, 4 fatty alcohol oxidases, and 6 alcohol dehydrogenases from the C. tropicalis genome. We also show that fatty acids with different chain lengths and degrees of unsaturation can be more efficiently oxidized by expressing different P450s within this strain background. Biocatalysis using engineered C. tropicalis is thus a potentially attractive biocatalytic platform for producing commodity chemicals from renewable resources.

Ophiocordyceps sinensis (Berk.) Sung et al. is popularly known as Yarsagumba (winter worm summer grass) in Nepal. It is a well-known entomogenous fungus distributed in alpine nival zone of trans-Himalayas and Tibetan Plateau. Its occurrence in extreme biological niche and cohabitation with insect [Thitarodes (Hepialus)] larvae has led to unique assemblage of metabolites including proteins and nitrogenous compounds, polysaccharides, sterols, nucleosides, fatty acids and their derivatives, vitamins and inorganics. In traditional Chinese medicine, it is one of the most trusted main ingredients for several preparations of remedy from wide range of human health conditions. Several biological activities from O. sinensis have been reported that include anti-inflammatory, antioxidant, antitumor, anti-metastatic, immunomodulatory, antimicrobial, insecticidal, hypolipidaemic, hypoglycemic, anti-ageing, lipolytic, neuroprotective, renoprotective effects, etc. The chemical constituents and their pharmacological uses are reviewed here highlighting the potentiality of this highly esteemed traditional Himalayan medicine.

For long-term growth of mammalian cells in perfused bioreactors, it is essential to monitor the concentration of dissolved oxygen (DO) present in the culture medium to ascertain the health of the cells. An optical oxygen sensor based on dynamic fluorescent quenching was developed for long-term continuous measurement of DO for NASA-designed rotating perfused bioreactors. Tris(4,7diphenyl-1,10-phenanthroline) ruthenium(II) chloride is employed as the fluorescent dye indicator. A pulsed, blue LED was chosen as the excitation light source. The sensor can be sterilized using an autoclave. The sensors were tested in a perfused rotating bioreactor supporting a BHK-21 (baby hamster kidney) cell culture over one 28-day, one 43-day, and one 180-day cell runs. The sensors were initially calibrated in sterile phosphate-buffered saline (PBS) against a blood-gas analyzer (BGA), and then used continuously during the entire cell culture without recalibration. In the 180-day cell run, two oxygen sensors were employed; one interfaced at the outlet of the bioreactor and the other at the inlet of the bioreactor. The DO concentrations determined by both sensors were compared with those sampled and measured regularly with the BGA reference. The sensor outputs were found to correlate well with the BGA data throughout the experiment using a single calibration, where the DO of the culture medium varied between 25 and 60 mm Hg at the bioreactor outlet and 80-116 mm Hg at the bioreactor inlet. During all 180 days of culture, the precision and the bias were F 5.1 mm Hg and À 3.8 mm Hg at the bioreactor outlet, and F 19 mm Hg and À 18 mm Hg at inlet. The sensor dynamic range is between 0 and 200 mm Hg and the response time is less than 1 minute. The resolution of the sensor is 0.1 mm Hg at 50 mm Hg, and 0.25 mm Hg at 130 mm Hg.

Small-scale roughness on bare agricultural soils determines very important properties such as infiltration, runoff or soil erosion. In this work, a method based on photogrammetric tools has been developed for the generation of automatic digital elevation models (DEMs) in agricultural soils, and is compared with a method based on very accurate offthe-shelf three-dimensional (3D) laser scanners. High resolution and accurate DEMs are generated by both of these non-contact techniques on two agricultural field plots of about 0.2 m 2 : (i) untilled and smooth soil, and (ii) very cloddy tilled surface. Furthermore, classical microrelief parameters such as Random Roughness and Maximum Depressional Storage are computed from grid DEMs obtained by laser scanning and close-range photogrammetry. Both techniques are compared on agricultural soils in terms of data acquisition and analysis performance, resolution, accuracy, and capability of representing microrelief and for computing microrelief parameters. Both the techniques tested have proved able to generate, under ideal conditions, DEMs with high accuracy (standard deviations of the residuals were 0.121 mm and 0.467 mm for laser and photogrammetry, respectively) and resolution (about 0.4 mm for laser scanning and 1 mm for the photogrammetric method). It is also worth noting that soil DEMs can be generated by both methods in less than 2 h, using devices in field assays that are not too expensive. Laser scanning produces a far more detailed surface allowing it to reproduce smaller aggregates than the photogrammetric method. On the other hand, data acquisition is faster with digital photogrammetry. The worse performance with either method happens when they are used on a rougher type of soil. Regarding the proposed non-contact methodologies, field soil microrelief can be attained in a fast, accurate, and economic way. In addition, the microrelief parameters computed from agricultural soil grid DEMs by laser scanning and photogrammetry show quite similar values. (M.A. Aguilar).

A monorail test system has been developed for studying the draught and power performance of narrow tillage tools operating at high speeds. The system is retrofitted to a small 10-m long linear soil bin, yet is capable of maintaining target tool speeds of 0.5-10 m s À1 over 1 to 3 m distances. Rigid vertical tools with three different blade shapes, two operating depths and two widths were tested in a silty clay loam with soft and hard compaction levels to demonstrate system capabilities. Soil disturbance was measured using a profilemeter. Results showed increases in power, tool draught and soil pulverization with increases in speed, and measurable effects of tool geometry (shape, width, operating depth) and of soil strength on power and draught-speed relationships.

Solid-state fermentation (SSF) has built up credibility in recent years in biotech industries due to its potential applications in the production of biologically active secondary metabolites, apart from feed, fuel, food, industrial chemicals and pharmaceutical products and has emerged as an attractive alternative to submerged fermentation. Bioremediation, bioleaching, biopulping, biobeneficiation, etc. are the major applications of SSF in bioprocesses which have set another milestone. Utilization of agro-industrial residues as substrates in SSF processes provides an alternative avenue and value-addition to these otherwise under-or non-utilized residues. Innovation is the key to success and it is imperative to be up-to-date with the changing demands of the industries and meet their needs for better product and services. Better understanding of biochemical engineering aspects, particularly on mathematical modeling and design of bioreactors (fermenters) has made it possible to scale-up SSF processes and some designs have been developed for commercialization, making the technology economically feasible. In future, SSF technology would be well developed at par with SmF if rationalization and standardization continues in current trend. This review describes the state-of-art scenario in totality on SSF although the focus is on the most recent developments of last 5 years or so on SSF processes and products developments.

Axillary shoots of Nicotiana benthamiana were regenerated from nodal explants in two weeks using MS media supplemented with the cytokinin, kinetin (0.5 mg/L), and the auxin, indole-3-butyric acid (IBA) (0.1 mg/L). Ninety two percent of shoots were 2.1-20 mm tall, a size ideal for root induction. After transfer to hormone-free MS they readily produced roots within seven days, with phenotypically normal, fully developed plants being obtained within four weeks. Leaf chlorosis due to iron deficiency was observed in plants over time, however, this was overcome by doubling the concentration of inorganic iron. This rapid micro-propagation system is particularly useful for the in vitro mass production of N. benthamiana plants for various biotechnological applications.

The Principal Investigator and his team would also like to highly appreciate the comments and valuable suggestions provided by all other stakeholders during consultation meetings and personal communication. Finally, the cooperation , active participation and valuable suggestions rendered to this study by other consultants also working for PAF activities are highly appreciated and acknowledged.

Agricultural fleet management is viewed as farmers' or machine contractors' decisionmaking concerning, for example, resource allocation, scheduling, routing, and real-time monitoring of vehicles and materials. In order to optimise this management task, fleet management tools are used for decision support to improve scheduling, routing and other operational measures for a fleet of agricultural machines. Additionally, fleet management involves the process of supervising the use and maintenance of machines and the associated administrative functions including the coordination and dissemination of tasks and related information for solving the heterogeneous scheduling and routing problems. In order to analyse the complex situation of how to develop an effective Fleet Management System, a conceptual model was developed based on a participatory approach and subsequent system analysis. This involved identification of the scope of the system, identification of user requirements, conceptual modelling, identification of actors and decision processes, and information-needs modelling. A holistic view and scope of the system is presented together with the system constraints. The results build on extracted information from selected actors in the agricultural domain. The systems components are depicted and linked to the subsequent conceptual model of the overall system as an outline for a subsequent tool development and implementation.

ßeta-galactosidaseenzyme is greatly dispersed in plant tissues and it's known to be implicated in hydrolyzing terminal non reducingß.D.Galactosyl residues from polysaccharides. This study involved the selection of the best extraction of the enzyme among seven methods.The phosphate buffer (pH7) had given a highest activity of crude enzyme was 8.85 U. The aqueous crude of almond seeds was used to detect of some compounds by phytochemical screening tests and using Thin layer chromatography(TLC). The protein content was concentrated and precipitated by ammonium sulphate (0-40)% as apartial purification. The purification stages of crude enzyme were achieved by using gel filtrationchromatography (sephadex-75) at a yield of (136.4)% with activity was 4.4 u/ml. followed by ion-exchange column chromatography (DEAE-cellulose A52) and five iso-enzymes were obtained at a yield of 93% with 4.6 times of purification fold for iso-enzyme (I) while 49.6% and 4.2 times of iso-enzyme (V).

The Magnetotactic Bacteria is a hot point of research field extending their applications frequently in the field of modern Science. Magnetosome Bacteria are unique in nature due to the presence of Magnetosomes. Magnetosome are now replacing the synthetically prepared Magnetic Nanoparticles using widely. In this research, The Magnetotactic Bacteria strain were isolated from the soil samples collected from Keetham Lake-Agra (India) and screened to optimised in-Vitro culture technique for magnetotactic Bacteria by utilising Hungate technique with most of the modification in growth medium and inoculation procedures.

Objective: The objective of the present study is to produce silver nanoparticles from marine fungi from south west coastal areas of Tamil Nadu and evaluating their potentials with special reference to anticancer activity. Methods: Hand core pushing technique is adapted to collect marine sediment samples along the coastal environs. Distinguished Aspergillus colonies were isolated and identified by wet mount procedure. The characterized Aspergillus consortium was subjected to produce silver nanoparticles. The extracellularly synthesized nanoparticles were characterized. Silver nanoparticles were evaluated for anticancer activity against MCF7 cell line by MTT assay. The IC50 values were determined. Results: Aspergillus consortium consist of A. niger, A. michelle and A. japonicus. Silver nanoparticles were extracellularly synthesized by the reduction of silver nitrate (AgNO3) to metallic silver (Ag +) ions results in the transformation of pale yellow to dark red colour. Constant shift was observed at 420 nm while monitoring the solution by UV-Vis spectrophotometer. The presence of important functional groups like – NH3 was confirmed by FT-IR Spectroscopy. Anticancer activity of the silver nanoparticles was evaluated against MCF7. There was 100% cell inhibition when concentration of the AgNPs reached 25 µg, 50 µg and 100 µg respectively in the test solution. Notably the IC50 value was found to be very lowest for the nanoparticles produced by A. japonicus and the value was found to be 1.47 µg/ml. Conclusions: Aspergillus consortium was found to be an ideal mycobiosystem for the production of silver nanoparticles with potential anticancer activity.

In recent years, considerable attention has been focused on the development of drug delivery system. There are number of reasons for the intense interest in new system. Recognition of the possibility of repeating successful drugs by applying the concepts and techniques of controlled release parenteral depot formulation drug delivery systems coupled with the increasing expense bringing new drug entities to market has encouraged the development of new drug delivery system. New systems are needed to deliver the novel, genetically engineered pharmaceuticals, i.e. peptides and proteins to their site of action without incurring significant immunogenic or biological inactivation. Treating enzyme deficient disease and cancer therapies can be improved by better targeting. Therapeutics efficacy and safety of drugs, administered by conventional methods, can be improved by more precise spatial and temporal placement within the body. Thereby reducing both size and number of doses. INTRODUCTION Primarily due to the steadily increasing number of biotechnology-derived drugs, parenteral depot formulations and their drug delivery technologies have received much attention in recent years. For many compounds which cannot be administered via the oral route, injectable or implantable depot formulations are presently the only viable option for reducing the frequency of administration, thereby increasing patient convenience and potentially improving compliance. Depot formulations have been developed for a broad range of applications, providing therapeutic effectiveness over periods ranging from one day to several years. A significant patient benefit, which should be the driving force for initiating the development of any depot formulation, not only requires that the dosing frequency can be substantially reduced in comparison with the corresponding standard formulation, but also that dosage form-related risks, such as dose dumping or poor carrier tolerability, are largely avoided. The Parenteral administration route is the most effective and common form of delivery for active drug substances with metabolic bio-availabilities drug for which the bio-availability in limited by high first pass metabolism effect of other physicochemical limitation and for drugs with a narrow therapeutic index. For this reason, whatever drug delivery technology that can reduce the total number of injection throughout the drug therapy period will be truly advantageous not only in terms of compliance, but also for potential to improve the quality of the therapy. Such reduction in frequency of drug dosing is achieved, in practice, by the use of specific formulation technologies that guarantee that the release of the active drug substance happens in a slow and predictable manner[1] .

Eight new primer sets were designed for PCR detection of (i) mono-oxygenase and dioxygenase gene sequences involved in initial attack of bacterial aerobic BTEX degradation and of (ii) catechol 2,3-dioxygenase gene sequences responsible for metacleavage of the aromatic ring. The new primer sets allowed detection of the corresponding genotypes in soil with a detection limit of 10 3 -10 4 or 10 5 -10 6 gene copies g À 1 soil, assuming one copy of the gene per cell. The primer sets were used in PCR to assess the distribution of the catabolic genes in BTEX degrading bacterial strains and DNA extracts isolated from soils sampled from different locations and depths (vadose, capillary fringe and saturated zone) within a BTEX contaminated site. In both soil DNA and the isolates, tmoA-, xylM-and xylE1-like genes were the most frequently recovered BTEX catabolic genes. xylM and 0167-7012/$ -see front matter D xylE1 were only recovered from material from the contaminated samples while tmoA was detected in material from both the contaminated and non-contaminated samples. The isolates, mainly obtained from the contaminated locations, belonged to the Actinobacteria or Proteobacteria (mainly Pseudomonas). The ability to degrade benzene was the most common BTEX degradation phenotype among them and its distribution was largely congruent with the distribution of the tmoA-like genotype. The presence of tmoA and xylM genes in phylogenetically distant strains indicated the occurrence of horizontal transfer of BTEX catabolic genes in the aquifer. Overall, these results show spatial variation in the composition of the BTEX degradation genes and hence in the type of BTEX degradation activity and pathway, at the examined site. They indicate that bacteria carrying specific pathways and primarily carrying tmoA/xylM/xylE1 genotypes, are being selected upon BTEX contamination. D